Zhang Shi-Yi, Lv Jian, Zhou Ze-Rui, Geng Peter X, Li Da-Wei, Qian Ruo-Can, Ju Huangxian
Key Laboratory for Advanced Materials, Feringa Nobel Prize Scientist Joint Research Center, Joint International Laboratory for Precision Chemistry, Frontiers Science Center for Materiobiology & Dynamic Chemistry, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, 200237, P. R. China.
Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing, 100871, P. R. China.
Adv Sci (Weinh). 2025 Feb;12(7):e2409344. doi: 10.1002/advs.202409344. Epub 2024 Dec 27.
Increased telomerase activity has been considered as a conspicuous sign of human cancers. The catalytic cores of telomerase involve a reverse transcriptase and the human telomerase RNA (hTR). However, current detection of telomerase is largely limited to its activity at the tissue and single-cell levels. To reveal the precise distribution of subcellular hTR and telomerase activity, here a modular engineered DNA nanodevice (DNA-ND) is designed capable of imaging hTR and telomerase activity in cytoplasm and nucleus, enabling colocalization analysis. DNA-ND is a modular DNA complex comprising hTR and telomerase activity detection modules, which respectively sense intercellular hTR and telomerase activity via target-sensitive allosteric transition of DNA switches, actuating orthogonal activation of fluorescence signals to achieve in situ co-imaging of hTR and telomerase activity. By integrating DNA-ND with specific localized signals, the DNA-ND based precise profiling of colocalization of hTR and telomerase activity in different cell lines as well as their dynamic changes during pharmacological interventions is demonstrated. Notably, the results suggest that the locations of hTR and telomerase activity are not exactly overlapped, indicating the influence of intracellular environment on the binding of hTR to telomerase.
端粒酶活性增加被认为是人类癌症的一个显著标志。端粒酶的催化核心包括一种逆转录酶和人类端粒酶RNA（hTR）。然而，目前对端粒酶的检测在很大程度上仅限于其在组织和单细胞水平的活性。为了揭示亚细胞hTR的精确分布和端粒酶活性，本文设计了一种模块化工程化DNA纳米器件（DNA-ND），能够对细胞质和细胞核中的hTR和端粒酶活性进行成像，从而实现共定位分析。DNA-ND是一种模块化DNA复合物，包含hTR和端粒酶活性检测模块，它们分别通过DNA开关的靶标敏感变构转变来感知细胞间的hTR和端粒酶活性，激活荧光信号的正交激活，以实现hTR和端粒酶活性的原位共成像。通过将DNA-ND与特定的定位信号整合，证明了基于DNA-ND的不同细胞系中hTR和端粒酶活性共定位的精确分析及其在药物干预过程中的动态变化。值得注意的是，结果表明hTR和端粒酶活性的位置并不完全重叠，这表明细胞内环境对hTR与端粒酶结合的影响。
